Tuyển tập báo cáo các nghiên cứu khoa học quốc tế ngành y học dành cho các bạn tham khảo đề tài: Pyrosequencing, a method approved to detect the two major EGFR mutations for anti EGFR therapy in NSCLC | Dufort et al. Journal of Experimental Clinical Cancer Research 2011 30 57 http content 30 1 57 Journal of Experimental Clinical Cancer Research RESEARCH Open Access Pyrosequencing a method approved to detect the two major EGFR mutations for anti EGFR therapy in NSCLC Sandrine Dufort1 2 Marie-Jeanne Richard1 2 Sylvie Lantuejoul2 3 and Florence de Fraipont1 2 Abstract Background Epidermal Growth Factor Receptor EGFR mutations especially in-frame deletions in exon 19 ALRE and a point mutation in exon 21 L858R predict gefitinib sensitivity in patients with non-small cell lung cancer. Several methods are currently described for their detection but the gold standard for tissue samples remains direct DNA sequencing which requires samples containing at least 50 of tumor cells. Methods We designed a pyrosequencing assay based on nested PCR for the characterization of theses mutations on formalin-fixed and paraffin-embedded tumor tissue. Results This method is highly specific and permits precise characterization of all the exon 19 deletions. Its sensitivity is higher than that of BigDye terminator sequencing and enabled detection of 3 additional mutations in the 58 NSCLC tested. The concordance between the two methods was very good . In the prospective analysis of 213 samples 7 samples were not analyzed and EGFR mutations were detected in 18 patients. However we observed a deficit of mutation detection when the samples were very poor in tumor cells. Conclusions pyrosequencing is then a highly accurate method for detecting ALRE and L858R EGFR mutations in patients with NSCLC when the samples contain at least 20 of tumor cells. Introduction Detection of mutations of the epidermal growth factor receptor EGFR gene is critical for predicting the response to therapy with tyrosine kinase inhibitors TKIs . gefitinib and erlotinib in patients with nonsmall-cell lung cancer NSCLC 1 . Practically all mutations are on exons 18 through 21 where they affect .
